Fig 7.

Addition of Mg²⁺ and deletion of lcrQ restore Yop translocation by the ΔrfaL mutant. Spleens from three healthy 6- to 8-week-old C57BL/6 mice were harvested and homogenized. Splenocytes were infected with wild-type (WT), ΔlcrQ, ΔrfaL, and ΔrfaL ΔlcrQ strains carrying YopM-Bla in RPMI (untreated) with or without EGTA or MgSO₄ at 37°C. Cells were stained with CCF2-AM and then analyzed by flow cytometry. The experiment was done in triplicate, and data are pooled from the three replicates, in order to determine the percentage of total splenocytes injected. Means and SEM are shown. Differences between strains were determined by one-way ANOVA and the Tukey post hoc test. ***, P < 0.001; **, P < 0.01.